BCMA CAR-T cells with enhanced activities

Abstract

Provided here are engineered immune cells that comprise a constitutively active chimeric cytokine receptor (CACCR) and a B-cell maturation antigen (BCMA) specific chimeric antigen receptor (CAR). Also provided herein are engineered immune cells that comprise one or more nucleic acids e.g. a bicistronic vector such as a viral vector that encode the CACCRs and BCMA CARs and engineered immune cells e.g. engineered autologous or allogeneic T cells that express both CACCRs and BCMA CARs from the nucleic acids. When present on chimeric antigen receptor (CAR)-bearing engineered immune cells, the CACCRs allow for increased immune cell activation, proliferation, persistence, and/or potency. Further provided herein are methods of making and using the engineered immune cells described herein, such as methods of treating a disease or condition by administering at least one appropriate dose of the cells to a patient suffering from the condition.

What is claimed is: 
     
       1. An engineered immune cell comprising an anti-B cell maturation antigen (BCMA) chimeric antigen receptor (CAR) and a constitutively active chimeric cytokine receptor (CACCR), wherein the CACCR comprises two monomers, each monomer comprising:
 a transmembrane domain; 
 a Janus Kinase (JAK)-binding domain; and 
 a recruiting domain comprising the amino acid sequence of SEQ ID NO: 78, 
 wherein the transmembrane domain and JAK-binding domain are present in a polypeptide (TM/JAK polypeptide) that comprises the amino acid sequence of SEQ ID NO: 12,
 wherein the CACCR does not comprise an extracellular ligand-binding domain, 
 wherein the anti-BCMA CAR comprises an extracellular ligand-binding domain, a transmembrane domain, and an intracellular signaling domain, 
 wherein the extracellular ligand-binding domain specifically recognizes and binds to human BCMA and comprises a single chain variable fragment (scFv), wherein the scFv comprises a heavy chain variable (VH) region and a light chain variable (VL) region; and 
 wherein the VH region comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 146, 147, or 148; a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 149 or 150; and a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 151; and the VL region comprises a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 152; a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 153; and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 154. 
 
 
     
     
       2. The engineered immune cell of  claim 1 , wherein the CACCR comprises the amino acid sequence of SEQ ID NO: 100, 181, or 185. 
     
     
       3. The engineered immune cell of  claim 1 , wherein the VH region comprises the amino acid sequence of SEQ ID NO: 144 and the VL region comprises the amino acid sequence of SEQ ID NO: 145. 
     
     
       4. The engineered immune cell of  claim 1 , wherein the intracellular signaling domain of the BCMA CAR comprises a CD3ζ (CD3zeta) signaling domain and/or a 4-1BB signaling domain. 
     
     
       5. The engineered immune cell of  claim 1 , wherein the BCMA CAR comprises a safety switch comprising the CD20 mimotope. 
     
     
       6. The engineered immune cell of  claim 1 , wherein the BCMA CAR comprises the amino acid sequence of SEQ ID NO: 140, 141 or 166. 
     
     
       7. An engineered immune cell that comprises one or more polynucleotides that encode a BCMA CAR polypeptide that comprises the amino acid sequence of SEQ ID NO: 140, 141 or 166 and a CACCR polypeptide that comprises the amino acid sequence of SEQ ID NO: 100, 181, or 185. 
     
     
       8. The engineered immune cell of  claim 7 , wherein a single polynucleotide encodes both the polypeptide that comprises the amino acid sequence of SEQ ID NO: 140, 141 or 166 and the polypeptide that comprises the amino acid sequence of SEQ ID NO: 100, 181, or 185. 
     
     
       9. An engineered immune cell of  claim 1  comprising a polynucleotide comprising the nucleic acid sequence of SEQ ID NO: 176. 
     
     
       10. The engineered immune cell of  claim 7 , wherein the one or more polynucleotides comprise a promoter. 
     
     
       11. The engineered immune cell of  claim 10 , wherein the promoter comprises an EF-1 alpha promoter. 
     
     
       12. The engineered immune cell of  claim 1 , wherein the engineered immune cell is an engineered T cell. 
     
     
       13. The engineered immune cell of  claim 1 , further comprising one or more genetic modifications in T cell receptor alpha constant region (TRAC) and/or CD52 gene to reduce or negate the expression of TCR alpha and/or CD52. 
     
     
       14. A method of treating a patient suffering from a disease or condition, wherein the method comprises administering an effective amount of the engineered immune cell of  claim 1  to the patient in need thereof. 
     
     
       15. The method of  claim 14 , wherein the disease or condition is multiple myeloma. 
     
     
       16. The method of  claim 14 , wherein the engineered immune cell is allogeneic to the patient. 
     
     
       17. The method of  claim 14 , wherein the method comprises administering at least one dose of the engineered immune cell to the patient, and further wherein one dose contains between about 7×10{circumflex over ( )}6 cells and about 480×10{circumflex over ( )}6 cells. 
     
     
       18. The method of  claim 17 , wherein the dose ranges from about 20×10{circumflex over ( )}6 cells/dose to about 480×10{circumflex over ( )}6 cells/dose or the at least one dose is about 20×10{circumflex over ( )}6 cells/dose, about 40×10{circumflex over ( )}6 cells/dose, about 80×10{circumflex over ( )}6 cells/dose, about 120×10{circumflex over ( )}6 cells/dose, about 240×10{circumflex over ( )}6 cells/dose, about 320×10{circumflex over ( )}6 cells/dose, about 360×10{circumflex over ( )}6 cells/dose, or about 480×10{circumflex over ( )}6 cells/dose. 
     
     
       19. One or more isolated polynucleotides encoding a BCMA CAR that comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 140, 141 or 166 and encoding a CACCR that comprises a polypeptide that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 100, 181, or 185; wherein the CACCR comprises the amino acid sequence of SEQ ID NO: 78 and SEQ ID NO: 12. 
     
     
       20. A vector that comprises the one or more isolated polynucleotides of  claim 19 . 
     
     
       21. The vector of  claim 20 , wherein the BCMA CAR comprises the amino acid sequence of SEQ ID NO: 140, 141 or 166 and the CACCR comprises the amino acid sequence of SEQ ID NO: 100, 181, or 185. 
     
     
       22. The vector of  claim 20 , wherein the vector further comprises a promoter. 
     
     
       23. The vector of  claim 22 , wherein the promoter comprises an EF-1 alpha promoter. 
     
     
       24. The vector of  claim 20 , wherein the vector is a lentiviral vector. 
     
     
       25. The vector of  claim 20  further comprising a mutant WPRE. 
     
     
       26. An engineered immune cell that comprises the one or more polynucleotides of  claim 19 . 
     
     
       27. A method of making an engineered immune cell of  claim 1  comprising introducing into a cell: at least one polynucleotide that encodes a BCMA CAR and at least one polynucleotide that encodes a CACCR, wherein the engineered immune cell expresses a BCMA CAR and a CACCR. 
     
     
       28. The method of  claim 27 , wherein the immune cell is a T cell, dendritic cell, killer dendritic cell, mast cell, NK-cell, macrophage, monocyte, B-cell or an immune cell derived from a stem cell. 
     
     
       29. The method of  claim 27 , wherein the cell is autologous. 
     
     
       30. The method of  claim 27 , wherein the cell is allogeneic. 
     
     
       31. The method of  claim 27 , wherein the polynucleotide further encodes at least one selectable marker. 
     
     
       32. The method of  claim 27 , wherein the polynucleotide is introduced into the cell by electroporation, transfection and/or viral transduction.